Figure_4.tif (797.33 kB)

Validation of NRL binding to corresponding peak regions by ChIP–qPCR.

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posted on 2012-04-12, 01:27 authored by Hong Hao, Douglas S. Kim, Bernward Klocke, Kory R. Johnson, Kairong Cui, Norimoto Gotoh, Chongzhi Zang, Janina Gregorski, Linn Gieser, Weiqun Peng, Yang Fann, Martin Seifert, Keji Zhao, Anand Swaroop

ChIP-qPCR was performed to validate NRL binding to 26 ChIP–Seq peak regions (left panel), and 5 non-peak regions (right panel) served as negative controls. The amount of ChIP DNA was measured by qPCR in triplicates using primers flanking the regions of interest. Normal IgG served as an antibody control when ChIP was performed using WT retinas (white bars). White bars (NRL Ab/IgG) represent fold change (FC) of qPCR signals comparing NRL ChIP DNA to the IgG control ChIP DNA. A separate set of ChIP assays was performed using NRL antibody to compare signals from WT retina to signals from Nrl^−/− retina (tissue control). Black bars (WT/Nrl^−/−) represent fold increase (Fc) of qPCR signals comparing NRL ChIP DNA from wild type C57BL/6 mouse retina to NRL ChIP DNA from Nrl^−/⁻ mouse retina. The ChIP-qPCR assays were performed twice. The representative results were shown as mean ± SD. P<0.01 for all by Student's t test.